Cephem compounds

ABSTRACT

The invention relates to intermediates useful in the preparation of antimicrobial compounds, the intermediates being of the formula: ##STR1## wherein R A  is --SCN or --S.sup.⊕ (R 3 ) 2 , wherein 
     R 3  is lower alkyl, and 
     R B  is carboxy or --COO.sup.⊖, 
     or a salt thereof, with the proviso that when R A  is --SCN, then R B  is carboxy and that when R A  is --S.sup.⊕ (R 3 ) 2 , then R B  is --COO.sup.⊕.

This application is a division of parent application Ser. No.: 785,712,filed Oct. 9, 1985, now U.S. Pat. No. 4,654,422, which in turn is adivision of application Ser. No. 511,237, filed July 6, 1983, now U.S.Pat. No. 4,563,449.

The present invention relates to new cephem compounds andpharmaceutically acceptable salts thereof. More particularly, it relatesto new cephem compounds and pharmaceutically acceptable salts thereof,which have antimicrobial activities and to processes for preparationthereof, to pharmaceutical composition comprising the same, and to amethod of using the same therapeutically in the treatment of infectiousdiseases in human being and animals.

Accordingly, it is one object of the present invention to provide newcephem compounds and pharmaceutically acceptable salts thereof, whichare active against a number of pathogenic microorganisms.

Another object of the present invention is to provide processes for thepreparation of new cephem compounds and pharmaceutically acceptablesalts thereof.

A further object of the present invention is to provide pharmaceuticalcomposition comprising, as active ingredients, said new cephem compoundsand pharmaceutically acceptable salts thereof.

Still further object of the present invention is to provide a method forthe treatment of infectious diseases caused by pathogenic bacteria inhuman being and animals.

The object new cephem compounds are novel and can be represented by thefollowing general formula (I). ##STR2## wherein R¹ is phosphono orprotected phosphono;

R² is lower aliphatic hydrocarbon group;

R³ is pyridinio or pyridiniothio, each of which may have suitablesubstituent(s), and

Y is N or CH.

According to the present invention, the new cephem compounds (I) can beprepared by processes which are illustrated in the following scheme.##STR3## wherein R¹, R², R³ and Y are each as defined above, R^(1a) is aprotected phosphono, R^(1b) is phosphono, R⁷ is amino or lower alkoxyand R⁸ is amino or morpholino.

Among the starting compounds of the present invention, the compound (II)is novel and can be prepared by the following methods. ##STR4## whereinR¹, R² and Y are each as defined above, R⁵ is an ester moiety of anesterified carboxy represented by a group of the formula: --COOR⁵,R^(1c) is dihalophosphoryl and R^(1d) is protected phosphono other thandihalophosphoryl.

Further, the compound (III) can be prepared by the following methods.##STR5## wherein Z is an acid residue, R⁶ is lower alkyl and R^(3a) ispyridinio which may have suitable substituent(s).

Regarding the object compounds (I), (Ia), (Ib), (Id) and (Ie) andstarting compounds (II), (IIa), (IV), (V), (VI), (VIa) and (VIb), it isto be understood that said object and starting compounds include synisomer, anti isomer and a mixture thereof. For example, with regard tothe object compound (I), syn isomer means one geometrical isomer havingthe partial structure represented by the following formula: ##STR6##(wherein R¹, R² and Y are each as defined above) and anti isomer meansthe other geometrical isomer having the partial structure represented bythe following formula: ##STR7## (wherein R¹, R² and Y are each asdefined above).

Regarding the other compounds as mentioned above, the syn isomer and theanti isomer can also be referred to the same geometrical isomers asillustrated for the compound (I).

Further, as to the object compounds (I) and (I_(b)), in case that R¹ ofthe compound (I) is phosphono [i.e. the same as the compound (I_(b))],said compounds (I_(b)) may also be alternatively represented by theformula: ##STR8## wherein R², R³ and Y are each as defined above, andboth of the compounds (I_(b)) and (I_(c)) are included within the scopeof the present invention.

In the present specification and claims, both of the compounds (I_(b))and (I_(c)) are represented by using one of the expressions therefor,that is by the formula: ##STR9## only for the convenient sake.

Suitable pharmaceutically acceptable salts of the object compounds (I)are conventional non-toxic salt and include a metal salt such as analkali metal salt (e.g. sodium salt, potassium salt, etc.) and analkaline earth metal salt (e.g. calcium salt, magnesium salt, etc.), anammonium salt, an organic base salt (e.g. trimethylamine salt,triethylamine salt, pyridine salt, picoline salt, dicyclohexylaminesalt, N,N'-dibenzylethylenediamine salt, etc.), and the like.

In the above and subsequent descriptions of the present specification,suitable examples and illustrations of the various definitions which thepresent invention include within the scope thereof are explained indetails as follows.

The term "lower" is intended to mean 1 to 6 carbon atoms, unlessotherwise indicated.

Suitable "protected phosphono" may include dihalophosphoryl (e.g.,dichlorophosphoryl, etc.); mono or di-esterified phosphono such asdi(lower)alkoxyphosphoryl(e.g. dimethoxyphosphoryl, diethoxyphosphoryl,dipropoxyphosphoryl, etc.), O-lower alkyl phosphono (e.g.O-methylphosphono, O-ethylphosphono, etc.) or the like; mono- ordi-amidated phosphono [e.g. diaminophosphoryl,(amino)(hydroxy)phosphoryl, etc.]; mono-esterified monoamidatedphosphono such as (lower alkoxy)(amino)phosphoryl [e.g.(methoxy)(amino)phosphoryl, (ethoxy)(amino)phosphoryl, etc.], (loweralkoxy)(morpholino)phosphoryl [e.g. (methoxy)(morpholino)phosphoryl,(ethoxy)(morpholino)phosphoryl, etc.] or the like; and the like.

Suitable lower aliphatic hydrocarbon group may include lower alkyl,lower alkenyl, lower alkynyl and the like.

Suitable "lower alkyl" is one having 1 to 6 carbon atom(s) and mayinclude methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl,pentyl, tert-pentyl, hexyl and the like, and preferably one having 1 to4 carbon atom(s).

Suitable "lower alkenyl" is one having 2 to 6 carbon atoms and mayinclude vinyl, allyl, isopropenyl, 1-propenyl, 2-butenyl, 3-pentenyl andthe like, and preferably one having 2 to 4 carbon atoms.

Suitable "lower alkynyl" is one having 2 to 6 carbon atoms and mayinclude ethynyl, 2-propynyl, 2-butynyl, 3-pentynyl, 3-hexynyl and thelike, and preferably one having 2 to 4 carbon atoms.

Suitable "substituent(s)" on "pyridinio" or "pyridiniothio" may includelower alkyl, halogen, lower alkoxy and the like.

Suitable "halogen" may be chlorine, bromine, iodine or fluorine.

Suitable "lower alkoxy" is one having 1 to 6 carbon atoms(s) and mayinclude methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy,pentyloxy, hexyloxy and the like, and preferably one having 1 to 4carbon atom(s).

Suitable ester moiety of an esterified carboxy represented by a group ofthe formula: --COOR⁵ may be the ones such as lower alkyl ester (e.g.methyl ester, ethyl ester, propyl ester, isopropyl ester, butyl ester,t-butyl ester, pentyl ester, t-pentyl ester, hexyl ester, etc.);

lower alkenyl ester (e.g. vinyl ester, allyl ester, etc.); lower alkynylester (e.g., ethynyl ester, propynyl ester, etc.); mono(or di ortri)-halo(lower)alkyl ester (e.g. 2-iodoethyl ester,2,2,2-trichloroethyl ester, etc.); ar(lower)alkyl ester, for example,phenyl(lower)alkyl ester which may be substituted with one or moresuitable substituent(s) (e.g. benzyl ester, 4-methoxybenzyl ester,4-nitrobenzyl ester, phenethyl ester, trityl ester, diphenylmethylester, bis(methoxyphenyl)methyl ester, 3,4-dimethoxybenzyl ester,4-hydroxy-3,5-ditertiarybutylbenzyl ester, etc.); or the like.

Suitable "acid residue" may include acyloxy, halogen as mentioned above,azido and the like.

Suitable "acyloxy" may include lower alkanoyloxy having 1 to 6 carbonatom(s) (e.g. formyloxy, acetoxy, propionyloxy, etc.) and the like, andpreferably one having 1 to 3 carbon atom(s).

Suitable "salt of HSCN" may include an alkali metal salt (e.g. sodiumsalt, potassium salt, etc.), salt with a heavy metal (e.g. cuprous salt,lead salt, etc.), ammonium salt, and the like.

Suitable "dihalophosphoryl" and "protected phosphono other thandihalophosphoryl" can be referred to the ones as exemplified for"protected phosphono".

Preferred embodiments of the object compound (I) are as follows.

Preferred embodiment of R¹ is phosphono, di(lower)alkoxyphosphoryl,O-lower alkyl phosphono, diaminophosphoryl, (amino)(hydroxy)phosphorylor (lower alkoxy)(morpholino)phosphoryl;

R² is lower alkyl or lower alkenyl;

R³ is a group of the formula: ##STR10## wherein X is hydrogen, halogenor lower alkoxy, or a group of the formula: ##STR11## wherein R⁴ islower alkyl; and Y is N or CH.

The processes for preparing the object compound (I) are explained indetails in the following.

Process 1

The object compound (I) or a salt thereof can be prepared by reactingthe compound (II) or its reactive derivative at the carboxy group or asalt thereof with the compound (III) or its reactive derivative at theamino group or a salt thereof.

Suitable reactive derivative at the amino group of the compound (III)may include conventional reactive derivative used in amidation, forexample, Schiff's base type imino or its tautomeric enamine type isomerformed by the reaction of the compound (III) with a carbonyl compound; asilyl derivative formed by the reaction of the compound (III) with asilyl compound such as bis(trimethylsilyl)acetamide,trimethylsilylacetamide or the like; a derivative formed by reaction ofthe compound (III) with phosphorus trichloride or phosgene, and thelike.

Suitable salt of the compound (III) may include an acid addition saltsuch as an organic acid salt (e.g. acetate, maleate, tartrate,benzenesulfonate, toluenesulfonate, etc.) or an inorganic acid salt(e.g. hydrochloride, hydrobromide, sulfate, phosphate, etc.); a metalsalt (e.g. sodium salt, potassium salt, calcium salt, magnesium salt,etc.); ammonium salt; an organic amine salt (e.g. triethylamine salt,dicyclohexylamine salt, etc.), and the like.

Suitable reactive derivative at the carboxy group of the compound (II)may include an acid halide, an acid anhydride, an activated amide, anactivated ester, and the like. The suitable example may be an acidchloride; an acid azide; a mixed acid anhydride with an acid such assubstituted phosphoric acid (e.g. dialkylphosphoric acid,phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoric acid,halogenated phosphoric acid, etc.), dialkylphosphorous acid, sulfurousacid, thiosulfuric acid, sulfuric acid, alkylcarbonic acid, aliphaticcarboxylic acid (e.g. pivalic acid, pentanoic acid, isopentanoic acid,

2-ethylbutyric acid, acetic acid or trichloroacetic acid, etc.),alkanesulfonic acid (e.g. methanesulfonic acid, ethanesulfonic acid,propanesulfonic acid, etc.) or aromatic carboxylic acid (e.g. benzoicacid, etc.); a symmetrical acid anhydride; an activated amide withimidazole, dimethylpyrazole, triazole or tetrazole; or an activatedester (e.g. cyanomethyl ester, methoxymethyl ester, dimethyliminomethyl[(CH₃)₂ N⁺ ═CH--] ester, vinyl ester, propargyl ester, p-nitrophenylester, 2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenylester, mesyl phenyl ester, phenylazophenyl ester, phenyl thioester,p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester,pyranyl ester, pyridyl ester, piperidyl ester, 8-quinolyl thioester, oran ester with N,N-dimethylhydroxylamine, 1-hydroxy-2-(1H)-pyridone,N-hydroxysuccinimide, N-hydroxyphthalimide or1-hydroxy-6-chloro-1H-benzotriazole, and the like. These reactivederivatives can be optionally selected from them according to the kindof the compound (II) to be used.

The salts of the compound (II) may be salts with an inorganic base suchas an alkali metal salts (e.g. sodium or potassium salt), or an alkalineearth metal salt (e.g. calcium or magnesium salt), a salt with anorganic base such as trimethylamine, triethylamine, pyridine, or thelike.

The reaction is usually carried out in a conventional solvent such aswater, acetone, dioxane, acetonitrile, chloroform, methylene chloride,ethylene chloride, tetrahydrofuran, ethyl acetate,N,N-dimethylformamide, N,N-dimethylacetamide, pyridine or any otherorganic solvent which does not adversely influence to the reaction.Among these solvents, hydrophilic solvents may be used in a mixture withwater.

When the compound (II) is used in free acid form or its salt form in thereaction, the reaction is preferably carried out in the presence of aconventional condensing agent such as N,N-dicyclohexylcarbodiimide;N-cyclohexyl-N'-morpholinoethylcarbodiimide;N-cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide;N,N-diethylcarbodiimide; N,N-diisopropylcarbodiimide;N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide;N,N-carbonylbis(2-methylimidazole);pentamethylene-ketene-N-cyclohexylimine;diphenylketene-N-cyclohexylimine; ethoxyacetylene; ethyl polyphosphate;isopropyl polyphosphate; diethyl phosphorochloridite; phosphorusoxychloride; phosphorus trichloride; phosphorus pentachloride; thionylchloride; oxalyl chloride; triphenylphosphine;N-ethyl-7-hydroxybenzisoxazolium fluoroborate;N-ethyl-5-phenylisoxazolium-3'-sulfonate;1-(p-chlorobenzenesulfonyloxy)-6-chloro-1H-benzotriazole; so-calledVilsmeier reagent, for example (chloromethylene)dimethylammoniumchloride produced by the reaction of dimethylformamide with thionylchloride or phosgene, a compound produced by the reaction ofdimethylformamide with phosphorus oxychloride, etc.; or the like.

The reaction may be also carried out in the presence of an inorganic oran organic base such as an alkali metal hydroxide, an alkali metalbicarbonate, alkali metal carbonate, alkali metal acetate,tri(lower)alkylamine, pyridine, N-(lower)alkylmorpholine,N,N-di(lower)alkylbenzylamine, N,N-di(lower)alkylaniline as exemplifiedbelow, or the like. When the base or the condensing agent is in liquid,it can be used also as a solvent. The reaction temperature is notcritical, and the reaction is usually carried out under cooling or atambient temperature.

In the present reaction, a syn-isomer of the object compound (I) can beobtained preferably by conducting the reaction of the compound (III)with a syn isomer of the starting compound (II), and "protectedphosphono group" for R¹, especially dihalophosphoryl, of the compound(II) may be converted into "phosphono" during the reaction orpost-treatment of the reaction to give the compound (I) wherein R¹ isphosphono, which is also included within the scope of the presentreaction.

PROCESS 2

The compound (Ib) or a salt thereof can be prepared by subjecting thecompound (Ia) or a salt thereof to elimination reaction of protectivegroup of phosphono.

Suitable salt of the compound (Ia) or (Ib) may be the ones asexemplified for the compound (I).

This elimination reaction can be conducted, for example, by reacting acompound (Ia) or a salt thereof with a trialkylsilyl halide (e.g.trimethylsilyl bromide, trimethylsilyl iodide, trimethylsilyl chloride,etc.), alkali metal halide (e.g. sodium iodide, potassium iodide, sodiumbromide, etc.), alkali metal thiocyanate (e.g. sodium thiocyanate,potassium thiocyanate, etc.), or the like.

The reaction is preferably carried out in a solvent such as methylenechloride, dimethylacetamide or any other organic ones which do notadversely influence the reaction. The reaction temperature is notcritical and the reaction is usually carried out under relatively mildconditions such as under cooling, at ambient temperature or slightlyelevated temperature.

PROCESS 3

The compound (Ie) or a salt thereof can be prepared by subjecting thecompound (Id) or a salt thereof to hydrolysis.

The present hydrolysis reaction may include a method using an acid andthe like.

Suitable acid may include an organic or an inorganic acid, for example,formic acid, sulfuric acid, trifluoroacetic acid, benzenesulfonic acid,nitric acid, p-toluenesulfonic acid, hydrochloric acid and the like, andpreferable acid is, for example, formic acid, trifluoroacetic acid,hydrochloric acid, etc. The acid suitable for the reaction can beselected according to the kind of group to be hydrolyzed.

The present reaction can be carried out in the presence or absence of asolvent. Suitable solvent may include a conventional organic solvent,water or a mixture thereof. When trifluoroacetic acid is used, theelimination reaction may preferably be carried out in the presence ofanisole.

The preparations for preparing the starting compound (II) are explainedin detail in the following.

PREPARATION A

The compound (V) or a salt thereof can be prepared by subjecting thecompound (IV) or its reactive derivative at the carboxy group or a saltthereof to esterification.

Suitable salt of the compound (IV) can be referred to the ones asexemplified for the compound (III), and salt of the compound (V) mayinclude acid addition salt as exemplified for the compound (III).

Suitable reactive derivative at the carboxy group of the compound (IV)can be referred to the ones as exemplified for the compound (II).

The esterifying agent to be used in the present esterification reactionmay include the compound represented by the formula:

    (R.sup.5).sub.2 SO.sub.4,

    R.sup.5a --N.sub.2

or

    R.sup.5 --X

wherein R⁵ is as defined above, R^(5a) is a group in which a hydrogen iseliminated from R⁵ and X is hydroxy or halogen.

Suitable halogen may include chlorine, bromine, iodine or fluorine.

The reaction using the esterifying agent represented by the formula:(R⁵)₂ SO₄ or R⁵ --X is usually carried out in a solvent such as water,acetone, methylene chloride, ethanol, ether, dimethylformamide or anyother solvent which does not adversely influence the reaction.

The present reaction is preferably carried out in the presence of a basesuch as an inorganic base or an organic base as aforementioned.

The reaction remperature is not critical and the reaction is usuallycarried out under cooling to heating around boiling point of thesolvent.

The reaction using the esterifying agent represented by the formula:R^(5a) --N₂ is usually carried out in a solvent such as ether,tetrahydrofuran or the like.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling or at ambient temperature.

PREPARATIONS B AND D

The compound (VI) or the compound (II) or its reactive derivative at thecarboxy group or a salt thereof can be prepared by subjecting thecompound (V) or a salt thereof or the compound (IV) or its reactivederivative at the carboxy group or a salt thereof to introductionreaction of phosphono or protected phosphono on amino, respectively.

Suitable reactive derivative at the carboxy group of the compound (IV)can be referred to the ones as exemplified for the compound (II).

Suitable agent to be used in the present introduction reaction mayinclude phosphorus halide (e.g. phosphorus trichloride, phosphoruspentachloride, etc.), phosphorus oxychloride and the like.

The present reaction is usually carried out in a solvent such asalkylene halide (e.g. methylene chloride, ethylene chloride, etc.),toluene or the like.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling, at ambient temperature or under warming.

In the present reaction, the reaction mixture obtained by reacting thecompound (IV) or (V) with the agent as above (e.g. phosphorus halide,etc.) wherein R¹ may be dihalophosphoryl may be further treated withwater to give the compound (II) or (VI) wherein R¹ is phosphono, and thesame reaction mixture may be further treated with alcohol such asalkanol (e.g. methanol, ethanol, etc.) or the like to give the compound(II) or (VI) wherein R¹ is esterified phosphono. Of course, the reactionproduct of the compound (II) or (VI) wherein R¹ is dihalophosphoryl canbe obtained from the reaction mixture as stated above by a conventionalisolation method and it can be used in the next step reaction.

The present reaction includes, within its scope, the case that thecarboxy group of the compound (IV) is converted into its reactivederivative during the reaction.

PREPARATION C

The compound (II) or a salt thereof can be prepared by subjecting thecompound (VI) to deesterification reaction.

Suitable salt of the compound (II) can be referred to the ones asexemplified for the compound (I).

The present reaction is carried out in accordance with a conventionalmethod such as hydrolysis, reduction or the like. Hydrolysis ispreferably carried out in the presence of a base or an acid. Suitablebase may include an inorganic base and an organic base such as an alkalimetal (e.g. sodium, potassium, etc.), an alkaline earth metal (e.g.magnesium, calcium etc.), the hydroxide or carbonate or bicarbonatethereof, trialkylamine (e.g. trimethylamine, triethylamine, etc.),picoline, 1,5-diazabicyclo[4,3,0]non-5-ene,1,4-diazabicyclo[2,2,2]octane, 1,8-diazabicyclo[5,4,0]undecene-7, or thelike. Suitable acid may include an organic acid (e.g. formic acid,acetic acid, propionic acid, trifluoroacetic acid, etc.) and aninorganic acid (e.g. hydrochloric acid, hydrobromic acid, sulfuric acid,etc.). Trifluoroacetic acid is preferably used in the presence of acation trapping agent (e.g. anisole, etc.).

The reaction is usually carried out in a solvent such as water,methylene chloride, tetrahydrofuran, an alcohol (e.g. methanol, ethanol,etc.), a mixture thereof or any other solvent which does not adverselyinfluence to the reaction. A liquid base or acid can be also used as thesolvent. The reaction temperature is not critical and the reaction isusually carried out under cooling to warming.

Reduction can be applied preferably for elimination of ester moiety suchas 4-nitrobenzyl, 2-iodoethyl, 2,2,2-trichloroethyl, or the like. Thereduction method applicable for the deesterification reaction mayinclude, for example, reduction by using a combination of a metal (e.g.zinc, zinc amalgam, etc.) or a salt of chrome compound (e.g. chromouschloride, chromous acetate, etc.) and an organic or inorganic acid (e.g.acetic acid, propionic acid, hydrochloric acid, etc.); and conventionalcatalytic reduction in the presence of a conventional metallic catalyst(e.g. palladium-carbon, etc.).

The preparations for preparing the starting compound (IIa) are explainedin detail in the following.

PREPARATION I

The compound (VIa) can be prepared by subjecting the compound (V) or asalt thereof to introduction reaction of dihalophosphoryl on amino.

The present reaction can be carried out according to similar manners tothose of Preparations B and D.

PREPARATION J

The compound (VIb) can be prepared by subjecting the compound (VIa) toconversion reaction of "dihalophosphoryl" to "protected phosphono otherthan dihalophosphoryl".

The present conversion reaction can be carried out by subjecting thecompound (VIa) to esterification and/or amidation.

The present esterification reaction can be conducted by reacting thecompound (VIa) with an alcohol.

Suitable alcohol may include an alkanol (e.g. methanol, ethanol,propanol, butanol, etc.) and the like.

The amidation reaction can be conducted by reacting the compound (VIa)with an amine.

Suitable amine may include ammonia, primary amine (e.g. methylamine,ethylamine, etc.), secondary amine (e.g. morpholin, dimethylamine, etc.)and the like.

The present esterification or amidation reaction is usually carried outin a solvent such as alkylene halide (e.g. methylene chloride, ethylenechloride, etc.), tetrahydrofuran, water or any other solvent which doesnot adversely affect the reaction.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling or at ambient temperature.

PREPARATION K

The compound (IIa) or a salt thereof can be prepared by subjecting thecompound (VIb) to deesterification reaction.

The present deesterification reaction can be carried out according to asimilar manner to that of Preparation C.

The Preparations for preparing the starting compound (IIIa) areexplained in detail in the following.

PREPARATIONS E AND G

The compound (IX) or a salt thereof or the compound (XII) or a saltthereof can be prepared by reacting the compound (VII) or a salt thereofwith the compound (VIII) or with the salt of the compound (XI),respectively.

Suitable salt of the compounds (VII), (IX) and (XII) can be referred tothe ones as exemplified for the compound (III).

The present reaction is usually carried out in a solvent such asalkanoic acid (e.g. acetic acid, trifluoroacetic acid, etc.),dimethylformamide, nitromethane, acetonitrile or any other solvent whichdoes not adversely affect the reaction.

The present reaction is preferably carried out in the presence ofsulfonic acid (e.g., trifluoromethanesulfonic acid, chlorosulfonic acid,etc.) and the like.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling to ambient temperature.

PREPARATIONS F AND H

The compounds (IIIa) or a salt thereof can be prepared by reacting thecompound (IX) or a salt thereof or the compound (XII) or a salt thereofwith the compound (X) or a salt thereof, respectively.

Suitable salt of the compound (IX) and (XII) can be referred to the onesexemplified for the compound (III).

Suitable salt of the compound (X) may include an acid addition salt asexemplified for the compound (III).

The present reaction may be carried out in a solvent such as water,phosphate buffer, acetone, chloroform, acetonitrile, nitrobenzene,methylene chloride, ethylene chloride, formamide, dimethylformamide,methanol, ethanol, ether, tetrahydrofuran, dimethylsulfoxide, or anyother organic solvent which does not adversely affect the reaction.Among the solvents, hydrophilic solvents may be used in a mixture withwater. The reaction is preferably carried out in around neutral medium.When the compound (IX) or (XII) is used in acid salt form, the reactionmay be conducted in the presence of a base, for example, inorganic basesuch as alkali metal hydroxide, alkali metal carbonate, alkali metalbicarbonate, organic base such as trialkylamine, and the like. Thereaction temperature is not critical, and the reaction is usuallycarried out under cooling, at ambient temperature, under warming orunder heating.

Thus obtained compounds according to Processes 1 to 3 as above may beconverted into pharmaceutically acceptable salts thereof by conventionalmanner.

The object compound (I) of the present invention exhibits highantimicrobial activity and inhibits the growth of a number ofmicroorganisms including pathogenic Gram-positive and Gram-negativebacteria.

For therapeutic administration, the cephalosporin compounds according tothe present invention are used in the form of pharmaceutical preparationwhich contain said compounds in admixture with a pharmaceuticallyacceptable carriers such as an organic or inorganic solid or liquidexcipient suitable for oral, parenteral or external administration. Thepharmaceutical preparations may be in solid form such as capsule,tablet, dragee, ointment or suppository, or in liquid form such assolution, suspension, or emulsion. If desired, there may be included inthe above preparations auxiliary substances, stabilizing agents, wettingor emulsifying agents, buffers and other commonly used additives.

While the dosage of the compounds may vary from and also depend upon theage and condition of the patient, an average single dose of about 50mg., 100 mg., 250 mg., and 500 mg. of the compounds according to thepresent invention may be effective for treating of infectious diseasescaused by a number of pathogenic bacteria. In general amounts, dailydose between 1 mg/body and about 1000 mg/body or even more may beadministered.

The object compound (I) of the present invention is characterized bypossessing higher solubility in water as compared with the correspondingfree aminothiadiazolyl or thiazolyl compound (i.e., R¹ --NH meansamino), which is slightly soluble in water, and by possessing a featurethat R¹ group can be split under physiological conditions to give thecorresponding free aminothiadiazolyl or thiazolyl compound.

Accordingly, the object of the present invention is to provide a moresoluble form of the corresponding aminothiadiazolyl or aminothiazolylcompound which is slightly soluble in water.

Now in order to show the utility of the object compounds (I), test dataon anti-microbial activity of a representative compound of the presentinvention are shown below.

TEST METHOD

One loopful of an overnight culture of each test strain inTrypticase-soy broth (10⁸ viable cells per ml.) was streaked on heartinfusion agar (HI-agar) containing graded concentrations of antibiotics,and the minimal inhibitory concentration (MIC) was expressed in terms ofμg/ml after incubation at 37° C. for 20 hours.

TEST COMPOUND

7-[2-Allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridinomethyl)-3-cephem-4-carboxylate(syn isomer)

TEST RESULTS

    ______________________________________                                        Test Bacteria M.I.C. (μg/ml)                                               ______________________________________                                        E. coli 31    0.78                                                            ______________________________________                                    

The following Preparations and Examples are given for the purpose ofillustrating the present invention.

PREPARATION 1

To a solution of 2-allyloxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)aceticacid (syn isomer) (10 g) in tetrahydrofuran (230 ml) was addedportionwise diphenyldiazomethane (12.76 g) under cooling in an ice bathand stirring and the mixture was stirred for 2 hours at roomtemperature. The resulting mixture was concentrated under reducedpressure until an crystallization began and stood overnight at roomtemperature. The precipitates were collected by filtration, washed withethyl acetate and dried to give diphenylmethyl2-allyloxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)acetate (syn isomer)(5.41 g), mp. 175° to 180° C. (dec.). The filtrate and the washings werecombined and evaporated to dryness. The residue was crystallized frommethylene chloride-diethyl ether (4:1) to give further crops of theobject compound (6.7 g), which was recrystallized from acetonitrile.

IR (Nujol): 3460, 1730, 1620, 1530, 1260, 1150, 1015 cm⁻¹.

NMR (DMSO-d₆, δ): 4.71 (2H, d, J=5 Hz), 5.05-5.50 (2H, m), 5.65-6.30(1H, m), 7.06 (1H, s), 7.40 (10H, s), 8.22 (2H, broad s).

PREPARATION 2

To a cold solution of phosphorus pentachloride (3.74 g) in methylenechloride (43 ml) was added diphenylmethyl2-allyloxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)acetate (syn isomer)(3.49 g) at -20° C. under stirring. To this mixture was added dropwisepyridine (2.37 g) under cooling below -22° C. in a dry ice-acetone bathand stirring, which was continued for one hour at -9° to -5° in anice-salt bath. After the mixture was cooled to -35° C. and pyridine(6.16 ml) in methylene chloride (36 ml) was added dropwise to themixture below -12° C. The mixture was stirred for 10 minutes at -10° to0° and for additional 10 minutes at 0° to 15° C. Cold water (70 ml) wasadded to the reaction mixture and the organic layer was separated out,washed with water (100 ml) and a saturated aqueous solution of sodiumchloride and then evaporated. The residual oil was subjected to columnchromatography on silica gel (140 g) using benzene/ethyl acetate(2/1-1/2) as eluents. The fractions containing the object compound werecombined and evaporated to give diphenylmethyl2-allyloxyimino-2-[5-dimethyoxyphosphorylamino-1,2,4-thiadiazol-3-yl]acetate(syn isomer) (2.90 g) as an oil.

It crystallized by allowing to stand in a refrigerator, mp. 91° to 95°C.

IR (Film): 3500, 3100-2800, 1750, 1590, 1530, 1450, 1390, 1280-1230,1185, 1115, 1070-1010 cm⁻¹.

NMR (DMSO-d₆, δ): 3.75 (6H, d, J=12 Hz), 4.75 (2H, d, J=5 Hz), 5.0-5.5(2H, m), 5.65-6.26 (1H, m), 7.07 (1H, s), 7.73 (10H, s)

Analysis for C₂₂ H₂₃ N₄ O₆ P; Calc'd.: C: 52.59, H: 4.61, N: 11.15.Found: C: 52.44, H: 4.83, N: 10.79.

PREPARATION 3

To a solution of diphenylmethyl2-allyloxyimino-2-[5-dimethoxyphosphorylamino-1,2,4-thiadiazol-3-yl]acetate(syn isomer) (4.0 g) and anisole (10 ml) in methylene chloride (10 ml)was added dropwise trifluoroacetic acid (22 ml) below -2° C. undercooling in an ice-salt bath and stirring, which was continued for 20minutes at -8° to -2° C. The reaction mixture was evaporated underreduced pressure and the residual oil was dissolved in a mixture ofethyl acetate (60 ml) and water (50 ml), adjusting to pH 6 with aqueoussodium bicarbonate. The aqueous layer was separated out and adjusted topH 1 with 6N-hydrochloric acid under an addition of ethyl acetate (60ml). The organic layer was separated out and the aqueous layer wasextracted with ethyl acetate. The organic layer and the extract werecombined, washed with a saturated aqueous solution of sodium chloride,dried over magnesium sulfate and evaporated to dryness. To the residuedissolved in ethyl acetate (8 ml) was added a solution of sodium acetate(0.98 g) in methanol (25 ml) and the mixture was evaporated to dryness.The residue was triturated in diisopropyl ether to give sodium2-allyloxyimino-2-[5-dimethoxyphosphorylamino-1,2,4-thiadiazol-3-yl]acetate(syn isomer) (2.67 g), mp. 131° to 137° C. (dec.).

IR (Nujol): 3420, 3100, 1690, 1670, 1615, 1530 cm⁻¹.

NMR (D₂ O, δ): 3.92 (6H, d, J=12 Hz), 4.80 (2H, m) 5.18 -5.70 (2H, m),5.70-6.48 (1H, m).

PREPARATION 4

The following compound was obtained according to a similar manner tothat of Preparation 1. Diphenylmethyl2-ethoxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)acetate (syn isomer), mp.193° to 196.5° C. (dec.).

IR (Nujol): 3460, 1735, 1610, 1530, 1500 cm⁻¹.

NMR (DMSO-d₆, δ): 1.20 (3H, t, J=7 Hz), 4.23 (2H, q, J=7 Hz), 7.10 (1H,s), 7.3 (10H, m), 8.27 (2H, s).

PREPARATION 5

The following compound was obtained according to a similar manner tothat of Preparation 2. Diphenylmethyl2-ethoxyimino-2-[5-dimethoxyphosphorylamine-1,2,4-thiadiazol-3-yl]acetate(syn isomer) mp. 128° to 130° C.

IR (Nujol): 3070, 1750, 1590, 1530, 1450, 1390, 1250, 1240, 1185, 1140,1110, 1095, 1060, 1040 cm⁻¹.

NMR (DMSO-d₆, δ): 1.18 (3H, t, J=7 Hz), 3.68 (6H, d, J=11 Hz), 4.20 (2H,q, J=7 Hz), 7.00 (1H, s), 7.30 (10H, s), 10.6-11.5 (1H, broad s).

PREPARATION 6

The following compound was obtained according to a similar manner tothat of Preparation 3. Sodium2-ethoxyimino-2-[5-dimethoxyphosphorylamino-1,2,4-thiadiazol-3-yl]acetate(syn isomer), mp. 142° to 148.5° C. (dec.).

IR (Nujol): 3500, 3430, 2700, 1670, 1610, 1530, 1400, 1280, 1270, 1180,1145, 1110, 1050, 1030 cm⁻¹.

NMR (D₂ O, δ): 1.34 (3H, t, J=7 Hz), 3.90 (6H, d, J=12 Hz), 4.31 (2H, q,J=7 Hz).

EXAMPLE 1

To a suspension of sodium2-allyloxyimino-2-[5-dimethoxyphosphorylamino-1,2,4-thiadiazol-3-yl]acetate(syn isomer) (358.5 mg) and sodium bicarbonate (42 mg) inN,N-dimethylacetamide (3.6 ml) was added dropwise methanesulfonylchloride (172 mg) under cooling in an ice bath and stirring, which wascontinued for 1.5 hours at the same temperature. On the other hand, amixture of 7-amino-3-(1-pyridiniomethyl)-3-cephem-4-carboxylatedihydrochloride (400 mg) and trimethylsilylacetamide (2 g) in methylenechloride (4 ml) was stirred at room temperature to give a solution andcooled to -30° C. The cold solution was added to the activated mixtureprepared above and the resulting mixture was stirred for 30 minutes at-20° to -15° and then for additional 20 minutes at -15° to 0° C. Themixture was poured into 6% aqueous solution of sodium bicarbonate (5ml), adjusted to pH 3 with 6N hydrochloric acid and evaporated to removemethylene chloride. The residual aqueous solution was diluted to 72 mlwith water and subjected to column chromatography on a nonionicadsorption resin "Diaion HP-20" (Trademark: prepared by MitsubishiChemical Industries) (36 ml). After the column was washed with water,the elution was carried out with 30% and 40% aqueous methanolsuccessively. The fractions containing the object compound werecollected, evaporated to remove methanol under reduced pressure andlyophilized to give7-[2-allyloxyimino-2-{5-dimethoxyphosphorylamino-1,2,4-thiadiazol-3-yl}acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (379 mg), mp. 157° to 165° C. (dec.).

IR (Nujol): 3350, 3200, 1770, 1670, 1610, 1520 cm⁻¹.

NMR (DMSO-d₆ +D₂ O, δ): 3.12 and 3.50 (2H, ABq, J=18 Hz) 3.67 (6H, d,J=11 Hz), 4.65 (2H, m), 5.0-6.0 (5H, m), 5.08 (1H, d, J=5 Hz), 5.72 (1H,d, J=5 Hz), 8.17 (2H, m), 8.53 (1H, m), 9.35 (2H, m).

EXAMPLE 2

To a solution of phosphorus pentachloride (4.99 g) in methylene chloride(60 ml) was added 2-ethoxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)acetylchloride monohydrochloride (syn isomer) (5.42 g) under cooling in an icebath and stirring, which was continued for one hour at room temperature.The mixture was evaporated to dryness and the residue was dissolved inacetone (30 ml). The solution was added dropwise to a solution of7-amino-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate dihydrochloride(5.42 g) in 40% aqueous acetone (50 ml) under cooling in an ice bath andstirring, adjusting to pH 6.5 with aqueous sodium bicarbonate during theaddition. After stirring for one hour in an ice bath, acetone wasremoved by evaporation and the remaining aqueous solution was adjustedto pH 1 with 6N hydrochloric acid. The resulting insoluble material wasremoved by filtration and the filtrate was subjected to columnchromatography on a non ionic adsorption resin "Diaion HP-20" (540 ml).

The elution was carried out with water, 5%, 10% and 20% aqueous methanolsuccessively. The fractions containing an object compound were collectedand concentrated to 10 ml under reduced pressure. The residual solutionwas poured into acetone (200 ml) under stirring and the resultingprecipitates were collected by filtration and dried to give7-[2-ethoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(2.59 g), mp. 165° to 173° C. (dec.).

IR (Nujol): 3200, 2350, 1780, 1670, 1630, 1510 cm⁻¹.

NMR (DMSO-d₆ +D₂ O, δ): 1.19 (3H, t, J=7 Hz), 3.1-3.7 (2H, m), 3.9-4.5(2H, m), 5.23 (1H, d, J=5 Hz), 5.0-6.1 (2H, m), 5.91 (1H, d, J=5 Hz),8.26 (2H, m), 8.65 (1H, m), 9.18 (2H, m).

EXAMPLE 3

To a solution of7-[2-allyloxyimino-2-{5-dimethoxyphosphorylamino-1,2,4-thiadiazol-3-yl}acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) in the reaction mixture prepared from sodium2-allyloxyimino-2-[5-dimethoxyphosphorylamino-1,2,4-thiadiazol-3-yl]acetate(syn isomer) (502 mg) and7-amino-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate dihydrochloride(560 mg) according to a similar procedure to that of Example 1 was addeddropwise trimethylsilyl bromide (3.22 g) under cooling in an ice bathand stirring, which was continued for one hour at room temperature. Tothe mixture was added acetic acid (0.5 ml) and the mixture was pouredinto diisopropyl ether (500 ml) under stirring. The resulting oilyproduct was separated out by decantation and dissolved in water (50 ml).The aqueous solution was subjected to column chromatography on a nonionic adsorption resin "Diaion HP-20" (50 ml). The elution was carriedout with water and 10% aqueous methanol. The fractions containing theobject compound were collected, evaporated to remove methanol underreduced pressure and lyophilized to give7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (250 mg), mp. 175° to 181° C. (dec.).

IR (Nujol): 3200, 2350, 1780, 1670, 1630, 1520 cm⁻¹.

NMR (DMSO-d₆ +D₂ O, δ): 3.51 (2H, m), 4.72 (2H, m), 5.06-6.30 (5H, m),5.23 (1H, d, J=5 Hz), 5.92 (1H, d, J=5 Hz), 8.27 (1H, m), 8.70 (1H, m),9.13 (2H, m).

EXAMPLE 4

The following compound was obtained according to similar manners tothose of Examples 1 to 3.7-[2-Ethoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-methyl-4-pyridiniothiomethyl)-3-cephem-4-carboxylate (syn isomer), mp.160° to 169° C. (dec.).

IR (Nujol): 3250, 1770, 1670, 1630, 1520 cm⁻¹.

NMR (DMSO-d₆ +D₂ O, δ): 1.24 (3H, t, J=7 Hz), 3.70 (2H, m), 4.22 (3H,s), 4.35 (4H, m), 5.21 (1H, d, J=5 Hz), 5.83 (1H, d, J=5 Hz), 7.95 (2H,d, J=7 Hz), 8.62 (2H, d, J=7 Hz).

PREPARATION 7

To a suspension of phosphorus pentachloride (243.36 g) in toluene (2.5l) was added 2-allyloxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)aceticacid (syn isomer) (102.6 g) under cooling at 0° C. and stirring, whichwas continued for 45 minutes at 4° to 8° C. The reaction mixture waspoured into ice-cold water (1.5 l) under stirring. The organic layer wasseparated out, washed with cold water (2 l) and brine (1 l), dried overmagnesium sulfate and evaporated to dryness. The residue was trituratedin diisopropyl ether (150 ml) and the mixture was stirred for 10 minutesat 3° C. The resulting precipitate was filtered, washed with colddiisopropyl ether (50 ml) and dried to give2-allyloxyimino-2-(5-dichlorophosphorylamino-1,2,4-thiadiazol-3-yl)acetylchloride (syn isomer) (128.7 g). mp. 128° to 129° C.

IR (Nujol): 1789, 1774, 1585, 1250, 1125, 1040, 985 cm⁻¹.

PREPARATION 8

The following compounds were obtained according to a similar manner tothat of Preparation 7.

(1)2-Ethoxyimino-2-(5-dichlorophosphorylamino-1,2,4-thiadiazol-3-yl)acetylchloride (syn isomer), mp. 115° C.

IR (Nujol): 1780, 1590, 1530, 1220, 1050, 960, 910 cm⁻¹.

NMR (d₆ -acetone, δ): 1.37 (3H, t, J=7 Hz), 4.45 (2H, q, J=7 Hz).

    ______________________________________                                        Analysis for C.sub.6 H.sub.6 N.sub.4 O.sub.3 PSCl.sub.3                       C             H      N         Cl   P                                         ______________________________________                                        calc'd: 20.48     1.71   15.93   30.33                                                                              8.82                                    found:  20.79     1.78   16.22   30.63                                                                              8.98                                    ______________________________________                                    

(2)2-Propoxyimino-2-(5-dichlorophosphorylamino-1,2,4-thiadiazol-3-yl)acetylchloride (syn isomer), mp. 127° to 130° C.

IR (Nujol): 1790, 1590, 1530, 1220, 1120, 1050, 1010, 940 cm⁻¹.

PREPARATION 9

To solution of phosphorus pentachloride (10.92 g) in methylene chloride(110 ml) was added2-allyloxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)acetic acid (synisomer) (11.4 g) under cooling at -20° C. and stirring, which wascontinued for 50 minutes at -15° to -5°. To the mixture was dropped asolution of methanol (2.4 g) in pyridine (15.82 g) at -30° to -10° understirring, which was continued for 20 minutes at -10° C. To the reactionmixture was dropped water (110 ml) and the mixture was adjusted to pH 2with 1N aqueous sodium hydroxide and then stirred for 30 minutes at 0°C. The resulting precipitate was filtered, washed with water and driedto give methyl 2-allyloxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (8.40 g).

mp. 167° to 168° C.

IR (Nujol): 3425, 3260, 3140, 1745, 1625, 1600, 1540, 1440, 1410, 1290,1145, 1075, 1015, 995 cm⁻¹.

NMR (d₆ -DMSO, δ): 3.83 (3H, s), 4.7 (2H, m), 5.0-5.5 (2H, m), 5.7-6.3(1H, m), 8.25 (2H, s).

PREPARTION 10

To a mixture of phosphorus pentachloride (65.4 g) and pyridine (37.3 g)in methylene chloride (654 ml) was added methyl2-allyloxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)acetate (syn isomer)(38.1 g) under cooling at -15° C. and stirring, which was continued for30 minutes at -10° to -5° C. To the reaction mixture was added water(300 ml) at --10° to 5° C. and the organic layer was separated out,washed with water, dried over magnesium sulfate and filtered. Thefiltrate was added to a solution of methanol (25.1 g) and pyridine (149g) in methylene chloride (785 ml) at -20° to -10° C. under stirring,which was continued for 30 minutes at -10° to -5° C. and 4 hours atambient temperature. The mixture was diluted with water (1 l) andadjusted to pH 1.0 with 6N hydrochloric acid. The organic layer wasseparated out, washed with water, dried over magnesium sulfate andevaporated to dryness. The residue was triturated in diethyl ether togive methyl2-allyloxyimino-2-(5-dimethoxyphosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (38.7 g). mp. 113° to 114° C.

IR (Nujol): 3080, 1745, 1595, 1535, 1430, 1390, 1270, 1250, 1230, 1110,1030 cm⁻¹.

NMR (d₆ -DMSO, δ): 3.73 (6H, d, J=12 Hz), 3.87 (3H, s), 4.7-5.0 (2H, m),5.1-5.5 (2H, m), 5.7-6.3 (1H, m).

PREPARATION 11

To a solution of methyl2-allyloxyimino-2-(5-dimethoxyphosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (37.3 g) in tetrahydrofuran (106 ml) was added 1N-aqueoussodium hydroxide (234 ml) and the mixture was stirred for 2 hours atambient temperature. The reaction mixture was adjusted to pH 5 with 6Nhydrochloric acid and washed with ethyl acetate. The aqueous layer wasseparated out, adjusted to pH 1.0 with 6N hydrochloric acid andextracted with ethyl acetate. The extract was washed with brine, driedover magnesium sulfate and evaporated. The residue dissolved in methanol(50 ml) was added dropwise to a solution of sodium acetate (8.7 g) inmethanol (90 ml) at ambient temperature under stirring, which wascontinued for one hour at the same temperature. To the mixture was addedethyl acetate (200 ml) and the resulting precipitate was collected byfiltration, washed with ethyl acetate and dried to give sodium2-allyloxyimino-2-(5-dimethoxyphosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (32.6 g), mp. 168° to 173° C. (dec.).

IR (Nujol): 3450, 1590, 1535, 1395, 1270, 1190, 1130, 1050, 1025, 945,845 cm⁻¹.

NMR (d₆ -DMSO, δ): 3.65 (6H, d, J=12 Hz),4.5-4.8 (2H, m) 5.0-5.6 (2H,m), 5.7-6.4 (1H, m).

PREPARATION 12

The following compound was obtained according to similar manners tothose of Preparations 2 and 10. Ethyl2-methoxyimino-2-(2-dimethoxyphosphorylaminothiazol-4-yl)acetate (synisomer), oil.

IR (Film): 3480, 3120, 3000, 2950, 2900, 1740, 1630, 1580, 1530, 1465,1450, 1375, 1290-1240, 1200-1170, 1170-1020, 970, 920, 850, 785 cm⁻¹.

NMR (DMSO-d₆, δ): 1.30 (3H, t, J=7 Hz), 3.72 (6H, d, J=12 Hz), 3.92 (3H,s), 4.33 (2H, q, J=7 Hz), 7.34 (1H, s), 10.29 (1, broad s).

PREPARATION 13

The following compound was obtained according to similar manners tothose of Preparations 3 and 11. Sodium2-methoxyimino-2-(2-dimethoxyphosphorylaminothiazol-4-yl)acetate (synisomer), mp. 152° to 160° C. (dec.).

IR (Nujol): 1620, 1545, 1400, 1275, 1185, 1045 cm⁻¹.

NMR (D₂ O, δ): 3.79 (6H, d, J=11 Hz), 3.91 (3H, s) 7.07 (1H, s).

PREPARATION 14

To a mixture of7-(2-thienylacetamido)-3-(3-chloro-1-pyridiniomethyl)-3-cephem-4-carboxylate(22.5 g) and N,N-dimethylaniline (60.0 g) in methylene chloride (400 ml)was added dropwise trimethylsilylchloride (50.0 g) at ambienttemperature under stirring, which was continued for 15 minutes at thesame temperature. The reaction mixture was cooled to -30° C. andphosphorus pentachloride (31.2 g) was added thereto under stirring,which was continued for one hour at -30° to -25° C. The reaction mixturewas added to a cold solution of 1,3-butanediol (45 g) in methylenechloride (400 ml) under cooling in an ice-bath and stirring, which wascontinued for one hour at ambient temperature. The resulting precipitatewas filtered, washed with methylene chloride and redissolved in methanol(50 ml). After the solution was treated with activated charcoal (1 g),the filtrate was poured into acetone (500 ml) under stirring, which wascontinued for 30 minutes at ambient temperature. The resultingprecipitate was filtered, washed with acetone and dried to give7-amino-3-(3-chloro-1-pyridiniomethyl)-3-cephem-4-carboxylatedihydrochloride (17.7 g), mp. 160° to 165° C. (dec.).

IR (Nujol): 3350, 1790, 1720, 1620, 1490, 1170 cm⁻¹.

NMR (D₂ O, δ): 3.50 and 3.80 (2H, ABq, J=18 Hz), 5.28 (1H, d, J=4 Hz),5.40 (1H, d, J=4 Hz), 5.48 and 5.80 (2H, ABq, J=14 Hz), 8.0-8.3 (1H, m)9.27-9.57 (3H, m).

PREPARATION 15

The following compound was obtained according to a similar manner tothat of Preparation 14.7-amino-3-(4-methoxy-1-pyridiniomethyl)-3-cephem-4-carboxylatehydrochloride

IR (Nujol): 3400, 1780, 1640, 1570, 1525, 1420 cm⁻¹.

NMR (D₂ O, δ): 3.20 and 3.57 (2H, ABq, J=18Hz), 4.10 (3H, s), 5.00 and5.20 (2H, ABq, J=14 Hz), 5.20 (1H, d, J=4 Hz), 5.23 (1H, d, J=4 Hz),7.47 (2H, d, J=7 Hz), 8.67 (2H, d, J=7 Hz).

PREPARATION 16

To a mixture of 7-aminocephalosporanic acid (6.0 g) and dimethyl sulfide(1.36 g) in acetonitrile (30 ml) was added trifluoromethanesulfonic acid(9.0 g) below 18° C. under cooling in an ice-bath and stirring, whichwas continued for 30 minutes at 15° to 18° C. To the reaction mixturewas added ethyl acetate (80 ml), and the mixture was seeded and stirredfor one hour at ambient temperature. A resulting precipitate wasfiltered, washed with ethyl acetate and dried to give7-amino-3-dimethylsulfoniomethyl-3-cephem-4-carboxylatebis(trifluoromethanesulfonate) (8.55 g), mp. 190° to 195° C. (dec.).

IR (Nujol): 3150, 3000, 1790, 1700, 1635, 1600, 1490, 1420, 1220, 1160,1020 cm⁻¹.

NMR (D₂ O, δ): 3.00 (6H, s), 3.67 and 3.87 (2H, ABq, J=18 Hz), 4.53 (2H,broad s), 5.20 (1H, d, J=4 Hz) 5.33 (1H, d, J=4 Hz).

    ______________________________________                                        Analysis for C.sub.12 H.sub.16 N.sub.2 O.sub.9 S.sub.4 F.sub.6                C             H      N         S    F                                         ______________________________________                                        calc'd: 25.09     2.81   4.88    22.32                                                                              19.84                                   found:  25.11     2.78   5.04    22.93                                                                              20.23                                   ______________________________________                                    

PREPARATION 17

To a solution of 7-amino-3-dimethylsulfoniomethyl-3 cephem-4-carboxylatebis(trifluoromethanesulfonate) (574.5 mg) in N,N-dimethylformamide (10ml) was added pyridine (395 mg) under cooling in an ice-bath andstirring, which was continued for 30 minutes at 0° to 5° C. The reactionmixture was diluted with water to 50 ml and the solution was subjectedto H.P.L.C. to identify the reaction product and calculate the yield. Itcontained 7-amino-3-(1-pyridiniomethy)-3-cephem-4-carboxylate (133.5mg).

PREPARATION 18

To a mixture of 7-aminocephalosporanic acid (45.0 g) and potassiumthiocyanate (14.55 g) in acetonitrile (225 ml) was added dropwisetrifluoromethanesulfonic acid (67.5 g) below 18° C. under cooling in anice-bath and stirring, which was continued for 30 minutes at 15° to 18°C. The reaction mixture containing7-amino-3-cyanothiomethyl-3-cephem-4-carboxylic acid was added to amixture of pyridine (118.5 g) and water (150 ml) at 30° to 33.5° C.under stirring. The resultant mixture was immediately poured into a coldmixture of isopropyl alcohol (900 ml) and diisopropyl ether (1.2 l)under cooling in an ice-bath and stirring. The resulting precipitate wasfiltered, washed with isopropyl alcohol and diisopropyl ether and thenredissolved in water (1.0 l). An insoluble material was filtered off andthe filtrate was passed through a column packed with acidic alumina (150g). The eluate (1.38 l) was concentrated to about 60 g of weight underreduced pressure and stirred for 30 minutes under cooling in anice-bath. The resulting precipitate was filtered, washed with methanoland diisopropyl ether and dried to give7-amino-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate thiocyanate (12.56g), mp. 175° to 180° C. (dec.).

IR (Nujol): 2600-2300, 2050, 1790, 1650, 1630, 1560, 1150, 1045 cm⁻¹.

NMR (D₂ O, δ): 3.33 and 3.67 (2H, ABq, J=18 Hz), 5.12 (1H, d, J=4 Hz),5.30 (1H, d, J=4 Hz), 5.33 and 5.51 (2H, ABq, J=14 Hz) 7.84-8.24 (2H,m), 8.4-8.7 (1H, m), 8.75-9.03 (2H, m).

PREPARATION 19

To a mixture of 7-aminocephalosporanic acid (40 g) and potassiumthiocyanate (15.7 g) in acetonitrile (200 ml) was added dropwisetrifluoromethanesulfonic acid (40 ml) below 15° C. under cooling in anice-bath and stirring, which was continued for 30 minutes at 5° to 15°C. and for 40 minutes at ambient temperature. The reaction mixture waspoured into cold water (400 ml) and stirred for 30 minutes. Theresulting precipitate was filtered, washed with water and acetone anddried to give 7-amino-3-cyanothiomethyl-3-cephem-4-carboxylic acid (26.2g), mp. 175° to 180° C. (dec.).

IR (Nujol): 3170, 2600, 2350, 2160, 1800, 1615, 1530 cm⁻¹.

NMR (D₂ O+DCl, δ): 3.80 (2H, s), 4.30 (2H, s), 5.20 (1H, d, J=5 Hz),5.40 (1H, d, J=5 Hz).

EXAMPLE 5

To a suspension of 7-amino-3-(1-pyridinomethyl)-3-cephem-4-carboxylatedihydrochloride dihydrate (56 g) in methylene chloride (1.12 l) wasadded trimethylsilylacetamide (280 g) and the mixture was stirred for 15minutes at ambient temperature. The solution was cooled at -20° C. and2-allyloxyimino-2-(5-dichlorophosphorylamino-1,2,4-thiadiazol-3-yl)acetylchloride (syn isomer) (51 g) was added thereto at the same temperatureunder stirring, which was continued for 20 minutes at -13° to -10° C.and for 30 minutes at -5° to 0° C. The reaction mixture was poured intoa saturated aqueous solution of sodium bicarbonate (1 l) under stirringand the aqueous layer was separated out. The organic layer was extractedtwice with water (800 ml, 400 ml) and the extracts were combined withthe aqueous layer. The combined aqueous solution was adjusted to pH 1.5with 6N hydrochloric acid and the resulting precipitate was filteredoff. The filtrate was subjected to column chromatography on nonionicadsorption resin "Diaion HP-20" (6 l). After the column was washed withwater (10 l), the elution was carried out with 20% aqueous methanol. Thefractions containing the object compound were combined, adjusted to pH6.0 with 1N aqueous sodium hydroxide and concentrated to a weight of 318g under reduced pressure. To the solution was added ion-exchange resin"Dowex 50 W 10 X (H+form)" (prepared by the Dow Chemical Co.) (120 g)and the mixture was stirred for 5 minutes. The resin was filtered off,washed with water (142 ml), and the filtrate and the washings werecombined. The combined solution was mixed with n-butanol (2.5 l) at -15°to -10° C. and to the resulting solution was added acetone (2.5 l)dropwise at -10° C. under stirring, which was continued for 40 minutesafter removing the cooling bath. The resulting precipitate was filtered,washed with acetone (500 ml) and dried to give7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (60.6 g), mp. 175° to 181° C. (dec.).

EXAMPLE 6

A suspension of crude7-amino-3-(1-methyl-4-pyridinothiomethyl)-3-cephem-4-carboxylate(purity: 59.9%, 14.68 g) in water (145 ml) was adjusted to pH 6.1 withtriethylamine and acetone (73 ml) was added thereto. To the solution wasadded portionwise2-ethoxyimino-2-(5-dichlorophosphorylamino-1,2,4-thiadiazol-3-yl)acetylchloride (syn isomer) (10.20 g) at 3° to 7° C. under cooling in anice-bath and stirring. The reaction mixture was kept at pH 6 to 7 withtriethylamine during the addition. After stirring for one hour at 3° to5° C., the reaction mixture was evaporated to remove acetone. Theaqueous solution was adjusted to pH 6.0 with aqueous sodium bicarbonate,stirred for 30 minutes at ambient temperature and then adjusted to pH1.5 with 6N hydrochloric acid. The resultant precipitate was filteredoff and the filtrate was subjected to column chromatography on anonionic adsorption resin "Diaion HP-20" (980 ml). After the column waswashed with water (4 l), the elution was carried out with aqueousmethanol (10%-30%). The eluates were combined, evaporated to removemethanol and lyophilized to give7-[2-ethoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-methyl-4-pyridiniothiomethyl)-3-cephem-4-carboxylate(syn isomer) (11.81 g), mp. 160° to 169° C. (dec.).

EXAMPLE 7

(a) To a mixture of sodium2-methoxyimino-2-(2-dimethoxyphosphorylaminothiazol-4-yl)acetate (synisomer) (3.20 g) and sodium bicarbonate (0.81 g) inN,N-dimethylacetamide (32 ml) was added methanesulfonyl chloride (1.88g) under cooling in an ice-bath and stirring, which was continued forone hour and cooled to -23° C. To the cold reaction mixture was added asolution of 7-amino-3-(1-pyridiniomethyl-3-cephem-4-carboxylatedihydrochloride dihydrate (3.52 g) and trimethylsilylacetamide (17.6 g)in methylene chloride (35 ml) at -23° to -20° C. under stirring, whichwas continued for 30 minutes at -20° to -12° C. and for 25 minutes at-12° to 3° C. to give a mixture containing7-[2-methoxyimino-2-(2-dimethoxyphosphorylaminothiazol-4-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer).

(b) To the mixture containing7-[2-methoxyimino-2-(2-dimethoxyphosphorylaminothiazol-4-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) was dropped trimethylsilyl bromide (14.80 g) at 3° to 8° C.under stirring, which was continued for 2.5 hours at ambienttemperature. The reaction mixture was poured into diisopropyl ether (1.5l) and the resultant resinous oil was separated by decantation. The oilwas dissolved in water (300 ml), adjusted to pH 1 with 1N hydrochloricacid and subjected to column chromatography on a nonionic adsorptionresin "Diaion HP-20" (320 ml). After the column was washed with water(1.4 l), the elution was carried out with 20% aqueous methanol. Thefractions containing the object compound were collected, evaporated toremove methanol and lyophilized to give7-[2-methoxyimino-2-(2-phosphonoaminothiazol-4-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (1.3 g), mp. 155° to 164° C. (dec.).

IR (Nujol): 3200, 1775, 1660, 1630, 1610, 1530, 1490, 1340, 1210, 1185,1155, 1060, 1040 cm⁻¹.

NMR (D₂ O+N_(a) HCO₃, δ): 3.25 and 3.70 (2H, ABq, J=18 Hz), 4.00 (3H,s), 5.31 (1H, d, J=5 Hz), 5.38 and 5.61 (2H, ABq, J=14 Hz), 5.88 (1H, d,J=5 Hz), 7.07 (1H, s), 8.19 (2H, m), 8.58 (1H, m), 8.98 (2H, m).

EXAMPLE 8

A suspension of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (5.81 g) in water (140 ml) was adjusted to pH 5.48 with7.4N aqueous ammonia and the solution was lyophilized to givemonoammonium salt of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (5.7 g), mp. 185° to 195° C. (dec.).

IR (Nujol): 3500-3100, 1770, 1673, 1605, 1535, 1287 cm⁻¹.

EXAMPLE 9

A suspension of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (5.81 g) in water (150 ml) was adjusted to pH 3.0 with 1Naqueous sodium hydroxide and the solution was lyophilized to givemonosodium salt of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (6.0 g), mp. 200° to 210° C. (dec.).

IR (Nujol): 3500-3100, 1770, 1670, 1630, 1610, 1525 cm⁻¹.

EXAMPLE 10

A suspension of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (5.81 g) in water (150 ml) was adjusted to pH 6.0 with 1Naqueous sodium hydroxide and the solution was lyophilized to givedisodium salt of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (6.1 g), mp. 205° to 215° C. (dec.).

IR (Nujol): 3500-3100, 1765, 1670, 1640-1600, 1530, 1290 cm⁻¹.

EXAMPLE 11

A suspension of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (5.81 g) in water (150 ml) was adjusted to pH 6.0 withcalcium hydroxide (760 mg). After a small amount of insoluble materialwas filtered off, the filtrate was lyophilized to give calcium salt of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (6.2 g), mp.>230° C.

IR (Nujol): 3500-3100, 1770, 1670, 1635, 1610, 1525, 1290 cm⁻¹.

EXAMPLE 12

7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (5.81 g) was reacted with potassium acetate to givedipotassium salt of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (6.40 g), mp. 143° to 148° C. (dec.).

IR (Nujol): 3150, 1760, 1660, 1605, 1520, 1280, 1150, 1010 cm⁻¹.

EXAMPLE 13

To a solution of monoammonium salt of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (500 mg) in water (2 ml) was added N,N-diethylacetamide(3.5 ml) and the mixture was allowed to stand for 3 days in arefrigerator. The resulting precipitate was collected by filtration,washed with 70% aqueous N,N-diethylacetamide and acetone successivelyand dried in air to give crystalline monoammonium salt of7-[2-allyloxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylatehexahydrates solvated with one molecule of N,N-diethylacetamide (synisomer) (200 mg), mp. 90° to 95° C.

IR (Nujol): 3400, 3150, 1774, 1680, 1614, 1575, 1538, 1338, 1315, 1290,1210 cm⁻¹.

NMR (D₂ O, δ): 1.12 (3H, t, J=7 Hz), 1.18 (3H, t, J=7 Hz), 2.10 (3H, s),3.20 and 3.70 (2H, ABq, J=18 Hz), 3.35 (2H, q, J=7 Hz), 3.42 (2H, q, J=7Hz). 4.8 (2H, m), 5.30 (1H, d, J=5 Hz), 5.93 (1H, d, J=5 Hz), 5.1-6.5(5H, m), 8.10 (2H, m), 8.60 (1H, m), 8.95 (2H, m).

    ______________________________________                                        Analysis for C.sub.26 H.sub.36 N.sub.9 O.sub.9 PS.sub.2.6H.sub.2 O                     C    H           N      H.sub.2 O                                    ______________________________________                                        calc'd:    38.00  5.89        15.34                                                                              13.15                                      found:     37.99  5.56        15.38                                                                              13.5                                       ______________________________________                                    

EXAMPLE 14

A solution of7-[2-ethoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-methyl-4-pyridiniothiomethyl)-3-cephem-4-carboxylate(syn isomer) (12.31 g) and sodium acetate (3.28 g) in water (60 ml) wasadded portionwise to acetone (1325 ml) at ambient temperature understirring. The resultant precipitate was collected by filtration, washedwith acetone and dried to give disodium salt of7-[2-ethoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-methyl-4-pyridiniothiomethyl)-3-cephem-4-carboxylate(syn isomer) (15.0 g), mp. 155° to 160° C. (dec.).

IR (Nujol): 3600-3100, 1760, 1675, 1605, 1530, 1285, 1225, 1115 cm⁻¹.

EXAMPLE 15

(a) A solution of7-[2-propoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (5.0 g) in water (50 ml) was adjusted to pH 4.2 with 1Nammonium hydroxide and lyophilized to give powdery monoammonium salt of7-[2-propoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (4.88 g).

(b) The monoammonium salt as obtained above (1.0 g) was dissolved inwater (4 ml) and N,N-diethylacetamide (12 ml) was added thereto. Themixture was allowed to stand at ambient temperature to precipitatecrystals and was further ice-cooled for one hour. The precipitatingcrystals were collected by filtration, in turn washed with a cold 80%aqueous solution of N,N-diethylacetamide (1.5 ml) and acetone and driedto give crystalline monoammonium salt of7-[2-propoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylatesolvated with one molecule of N,N-diethylacetamide (syn isomer) (950mg), mp. 80° to 85° C.

IR (Nujol): 3400, 3200, 1770, 1680, 1620, 1540, 1490, 1340, 1210, 1150,1070, 1050, 1020, 1000, 930 cm⁻¹.

NMR (D₂ O, δ): 0.90 (3H, t, J=7 Hz), 1.08 (3H, t, J=7 Hz), 1.17 (3H, t,J=7 Hz), 1.53-1.98 (2H, m). 2.10 (3H, s), 3.30 (2H, t, J=7 Hz), 3.33(2H, t, J=7 Hz), 3.17 and 3.70 (2H, ABq, J=18 Hz), 4.27 (2H, t, J=7 Hz),5.30 (1H, d, J=5 Hz), 5.30 and 5.63 (2H, ABq, J=14 Hz), 5.90 (1H, d, J=5Hz), 8.10 (2H, m), 8.58 (1H, m), 8.97 (2H, m).

EXAMPLE 16

The following compounds were obtained according to similar manners tothose of aforesaid Examples.

(1)7-[2-Propoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer), mp. 177° to 182° C. (dec.).

IR (Nujol): 3200, 1780, 1650, 1630, 1520, 1485 cm⁻¹.

NMR (D₂ O, δ): 0.90 (3H, t, J=7 Hz), 1.47-2.0 (2H, m), 3.23 and 3.77(2H, ABq, J=18 Hz), 4.27 (2H, t, J=7 Hz), 5.33 (1H, d, J=5 Hz), 5.37 and5.77 (2H, ABq, J=14 Hz), 5.93 (1H, d, J=5 Hz), 8.10 (2H, m), 8.58 (1H,m), 8.95 (2H, m).

(2)7-[2-Ethoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(4-methoxy-1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer), mp. 165° to 170° C. (dec.).

IR (Nujol): 3200, 1780, 1670, 1640, 1570, 1520 cm⁻¹.

NMR (D₂ O, δ): 1.30 (3H, t, J=7 Hz), 3.30 and 3.63 (2H, ABq, J=18 Hz),4.10 (3H, s), 4.33 (2H, q, J=7 Hz), 5.20 and 5.47 (2H, ABq, J=14 Hz),5.28 (1H, d, J=5 Hz), 5.88 (1H, d, J=5 Hz), 7.43 (2H, d, J=7 Hz), 8.65(2H, d, J=7 Hz).

(3)7-[2-Ethoxyimino-2-(5-phosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(3-chloro-1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer), mp. 170° to 175° C. (dec.).

IR (Nujol): 3200, 2300, 1775, 1670, 1630, 1510 cm⁻¹.

NMR (D₂ O, δ): 1.30 (3H, t, J=7 Hz), 3.35 and 3.73 (2H, ABq, J=14 Hz),4.33 (2H, q, J=7 Hz), 5.32 (1H, d, J=5 Hz), 5.43 and 5.78 (2H, ABq, J=14Hz). 5.93 (1H, d, J=5 Hz), 7.93-8.27 (1H, m), 8.53-8.80 (1H, m),8.87-9.08 (1H, m), 9.17 (1H, broad s).

PREPARATION 20

Phosphorus pentachloride (21.98 g) was dissolved in methylene chloride(270 ml) at room temperature and cooled at -20° C. To the mixture weresuccessively added methyl2-ethoxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)acetate (syn isomer)(13.5 g) and pyridine (13.91 g) under stirring and keeping below -10° C.The mixture was stirred for 30 minutes and poured into 1N aqueoushydrochloric acid (176 ml). The organic layer was separated out, washedwith a saturated aqueous solution of sodium chloride, dried andevaporated under reduced pressure. The resulting precipitates werecollected by filtration, washed with diisopropyl ether and dried to givemethyl2-ethoxyimino-2-(5-dichlorophosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (6.93 g). The crude product was used to the followingreaction without further purification.

PREPARATION 21

To a cold solution of methyl2-ethoxyimino-2-(5-dichlorophosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (600 mg) in tetrahydrofuran was added aqueous conc.ammonia(0.48 ml) and the mixture was stirred for 30 minutes under cooling in anice-bath. The reaction mixture was evaporated, diluted with an aqueoussolution of sodium chloride (20 ml) and extracted with ethyl acetate.The extract was dried and evaporated. The residue was triturated in amixed solvent of tetrahydrofuran and diisopropyl ether and the resultingprecipitates were filtered, washed with diisopropyl ether to give methyl2-ethoxyimino-2-(5-diaminophosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (418 mg). The crude product was recrystallized fromacetonitrile, mp 191° to 193° C. (dec.).

IR (Nujol): 3200, 3130, 1755, 1515, 1275, 1230 cm⁻¹.

NMR (DMSO-d₆ +D₂ O, δ): 1.21 (3H, t, J=7 Hz), 3.80 (3H, s), 4.22 (2H, q,J=7 Hz).

Anal. Calcd. for C₇ H₁₃ N₆ O₄ PS: C, 27.28; H, 4.25; N, 27.26. Found: C,26.88; H, 4.12; N, 26.57.

PREPARATION 22

To a suspension of phosphorus pentachloride (48.59 g) in methylenechloride (630 ml) were added methyl2-allyloxyimino-2-(5-amino-1,2,4-thiadiazol-3-yl)acetate (syn isomer)(31.44 g) at -25° C. and then pyridine (30.76 g) at -20° to -11° C.under stirring, which was continued for 30 minutes at -10° to -3° C. Thereaction mixture was poured into a mixture of 1N aqueous hydrochloricacid (390 ml) and crushed ice. The organic layer was separated out,washed with a saturated aqueous solution of sodium chloride, dried andevaporated to dryness. The residue was triturated in diisopropyl etherto give methyl2-allyloxyimino-2-(5-dichlorophosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (29.02 g). The crude product was used to the followingreaction without further purification.

PREPARATION 23

To a solution of methyl2-allyloxyimino-2-(5-dichlorophosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (359 mg) in methylene chloride (3.6 ml) were added pyridine(79 mg) at -9° C. and then a solution of ethanol (46 mg) in methylenechloride (2 ml). The mixture was stirred for 30 minutes at -11° to -10°C., for 25 minutes at -10° to 0° C. and for one hour and 40 minutes atroom temperature. The reaction mixture was chilled in an ice-bath andmorpholine (174 mg) was added thereto under stirring, which wascontinued for 50 minutes at room temperature. Water (5 ml) was added tothe mixture. The organic layer was separated out, washed with asaturated aqueous solution of sodium chloride, dried and evaporated todryness. The residue was subjected to column chromatography on silicagel (10 g). The elution was carried out with a mixed solvent (ethylacetate/chloroform=2/1). The fractions containing the object compoundwere collected and evaporated to dryness to give methyl2-allyloxyimino-2-[5-(ethoxy)(morpholino)phosphorylamino-1,2,4-thiadiazol-3-yl]acetate(syn isomer) (137 mg), oil.

IR (Nujol): 3100, 3000, 1750, 1530, 1390 cm⁻¹.

NMR (CDCl₃, δ): 1.37 (3H, t, J=7 Hz), 3.20 (4H, m), 3.67 (4H, m), 3.96(3H, s), 4.18 (2H, m), 4.78 (2H, d, J=5 Hz), 5.43-5.10 (2H, m),5.78-6.20 (1H, m).

PREPARATION 24

To a solution of methyl2-allyloxyimino-2-(5-dichlorophosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (5.0 g) in methylene chloride (100 ml) were successivelyadded pyridine (3.3 g) and a solution of ethanol in methylene chloride(100 ml) under cooling at -35° C. to -25° C. and stirring. The mixturewas stirred for 20 minutes at -20° to -10° C., for 30 minutes at 5° C.and then for two hours at room temperature, and poured into water (100ml). The organic layer was separated, washed with 1N aqueoushydrochloric acid and a saturated aqueous solution of sodium chloride,dried over magnesium sulfate and evaporated to dryness under reducedpressure to give methyl2-allyloxyimino-2-(5-diethoxyphosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (5.25 g) as yellow oil.

IR (Film): 3500, 3100, 3000, 2900, 2800, 1750, 1600, 1530, 1510 cm⁻¹.

NMR (CDCl₃, δ): 1.37 (6H, t, J=7 Hz), 3.95 (3H, s), 4.22 (4H, m),4.68-4.90 (2H, m), 5.07-5.55 (2H, m), 5.67-6.37 (1H, m), 6.85-7.40 (1H,m).

PREPARATION 25

The following compounds were obtained according to similar manners tothose of Preparation 3 and 11.

(1) Sodium2-ethoxyimino-2-(5-diaminophosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer), mp 194° to 197° C. (dec.).

IR (Nujol): 3250, 1610, 1530, 1400, 1200 cm⁻¹.

NMR (DMSO-d₆ +D₂ O, δ): 1.20 (3H, t, J=7 Hz), 4.04 (2H, q, J=7 Hz).

(2) Sodium2-allyloxyimino-2-[5-(ethoxy)(morpholino)phosphorylamino-1,2,4-thiadiazol-3-yl]acetate(syn isomer), mp 172° to 177° C. (dec.).

IR (Nujol): 3400, 1620, 1530, 1265, 1115 cm⁻¹.

NMR (DMSO-d₆, δ): 1.34 (3H, t, J=7 Hz), 3.17 (4H, m), 3.60 (4H, m), 4.12(2H, m), 4.63 (2H, d, J=5 Hz), 4.92-5.63 (2H, m), 5.67-6.35 (1H, m).

(3) Sodium2-allyloxyimino-2-(5-diethoxyphosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer), mp 156° to 162° C. (dec.).

IR (Nujol): 1710, 1610, 1535, 1400 cm⁻¹.

NMR (DMSO-d₆ +D₂ O, δ): 1.22 (6H, t, J=7 Hz), 4.02 (4H, m), 4.55 (2H, d,J=5 Hz), 5.00-5.50 (2H, m), 5.67-6.25 (1H, m).

EXAMPLE 17

To a mixture of sodium2-ethoxyimino-2-(5-diaminophosphorylamino-1,2,4-thiadiazol-3-yl)acetate(syn isomer) (2.10 g) and sodium bicarbonate (1.12 g) inN,N-dimethylacetamide (21 ml) was added methane-sulfonyl chloride (1.03ml) under cooling in an ice-bath and stirring, which was continued forone hour and 45 minutes and cooled to -20° C. To the cold reactionmixture was added a solution of7-amino-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate dihydrochloridedihydrate (2.39 g) and monotrimethylsilylacetamide (11.96 g) inmethylene chloride (48 ml) at -18° C. under stirring, which wascontinued for 30 minutes at -18° to -12° C. and for 30 minutes at -10°to 0° C. The reaction mixture was poured into diisopropyl ether (700 ml)and the resulting precipitates were separated by decantation. Theresidue was dissolved in water (100 ml), adjusted to pH 3.8 with 1Nhydrochloric acid and subjected to column chromatography on a non-ionicadsorption resin "Diaion HP-20" (105 ml). After the column was washedwith water (500 ml), the elution was carried out with 20% aqueousmethanol. The fractions containing the object compound were collected,evaporated to remove methanol and lyophilized to give7-[2-ethoxyimino-2-(5-diaminophosphorylamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (1.11 g), mp 123° to 132° C. (dec.).

IR (Nujol): 3200, 1770, 1660, 1610, 1510 cm⁻¹.

NMR (DMSO-d₆ +D₂ O, δ): 1.25 (3H, t, J=7 Hz), 3.12, 3.55 (2H, ABq, J=17Hz), 4.19 (2H, q, J=7 Hz), 5.10 (1H, d, J=5 Hz), 5.23, 5.67 (2H, ABq,J=14 Hz), 5.75 (1H, d, J=5 Hz), 8.17 (2H, m), 8.63 (1H, m), 9.41 (2H, d,J=7 Hz).

EXAMPLE 18

The following compounds were obtained according to similar manners tothose of aforesaid Examples.

(1)7-[2-Allyloxyimino-2-{5-(ethoxy)(morpholino)phosphorylamino-1,2,4-thiadiazol-3-yl}acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer), mp 132° to 141° C. (dec.).

IR (Nujol): 3400, 3200, 1770, 1670, 1610, 1510 cm⁻¹.

NMR (DMSO-d₆ +D₂ O, δ): 1.22 (3H, t, J=8 Hz), 3.00 (4H, m), 3.46 (4H,m), 3.99 (2H, m), 4.59 (2H, d, J=5 Hz), 4.90-6.20 (4H, m), 5.00 (1H, d,J=5 Hz), 5.66 (1H, d, J=5 Hz), 8.06 (2H, m), 8.50 (1H, m), 9.26 (2H, d,J=5 Hz).

(2)7-[2-Ethoxyimino-2-{5-(amino)(hydroxy)phosphorylamino-1,2,4-thiadiazol-3-yl}acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer), mp 150` to 157° C. (dec.).

IR (Nujol): 3200, 1775, 1660, 1630, 1520 cm⁻¹.

(3)7-[2-Allyloxyimino-2-(5-O-ethylphosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer), mp 157° to 164° C. (dec.).

IR (Nujol): 3200, 1780, 1670, 1630, 1510 cm⁻¹.

(4)7-[2-Allyloxyimino-2-(5-diethoxyphosphorylamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer), mp 124° to 131° C. (dec.).

IR (Nujol): 3400, 3200, 1775, 1670, 1610, 1520 cm⁻¹.

NMR (D₂ O, δ): 1.38 (6H, t, J=7 Hz), 3.27, 3.70 (2H, ABq, J=18 Hz), 4.29(4H, m), 4.87 (2H, m), 5.10-5.70 (4H, m), 5.77-6.16 (1H, m), 5.33 (1H,d, J=5 Hz), 5.95 (1H, d, J=5 Hz), 8.16 (2H, m), 8.59 (1H, m), 9.05 (2H,m).

EXAMPLE 19

To a cold aqueous 1N hydrochloric acid was added7-[2-ethoxyimino-2-(5-diaminophosphorylamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (1.0 g) and the mixture was stirred for seven minutes in anice-bath. The reaction mixture was adjusted to pH 2 with aqueous sodiumbicarbonate and subjected to column chromatography on a non-ionicadsorption resin "Diaion HP-20" (100 ml). After the colunn was washedwith water (400 ml), the elution was carried out with 10% aqueousmethanol. The fractions containing the object compound were collected,concentrated to about 10 ml under reduced pressure and poured intoacetone (100 ml). The resulting precipitates were collected byfiltration, washed with acetone and dried to give7-[2-ethoxyimino-2-{5-(amino)(hydroxy)phosphorylamino-1,2,4-thiadiazol-3-yl}acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (0.37 g), mp 150° to 157° C. (dec.).

IR (Nujol): 3200, 1775, 1660, 1630, 1520 cm⁻¹.

NMR (DMSO-d₆ +D₂ O, δ): 1.19 (3H, t, J=7 Hz), 3.00-3.80 (2H, m), 4.12(2H, q, J=7 Hz), 5.10-5.80 (2H, m), 5.77 (1H, d, J=5 Hz), 8.18 (2H, m),8.55 (2H, m), 9.23 (2H, m).

EXAMPLE 20

A solution of7-[2-allyloxyimino-2-{5-(ethoxy)(morpholino)phosphorylamino-1,2,4-thiadiazol-3-yl}acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (2.80 g) in 1N aqueous hydrochloric acid (28 ml) wasstirred for 4 hours at 40° C. The reaction mixture was diluted withwater (70 ml) and subjected to column chromatography on non-ionicadsorption resin "Diaion HP-20" (112 ml). After the column was washedwith water (500 ml), the elution was carried out with 30% aqueousmethanol. The fractions containing the object compound were combined,concentrated to about 20 ml under reduced pressure. The residue waspoured to acetone (300 ml) and the resulting precipitates were filtered,washed with acetone and dried to give7-[2-allyloxyimino-2-(5-O-ethylphosphonoamino-1,2,4-thiadiazol-3-yl)acetamido]-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate(syn isomer) (1.16 g), mp 157° to 164° C. (dec.).

IR (Nujol): 3200, 1780, 1670, 1630, 1510 cm⁻¹.

NMR (D₂ O, δ): 1.22 (3H, t, J=7 Hz), 3.30, 3.72 (2H, ABq, J=19 Hz), 3.96(2H, m), 5.10-6.20 (5H, m), 5.22 (1H, d, J=5 Hz), 5.93 (1H, d, J=5 Hz),8.10 (2H, m), 8.61 (1H, m), 8.97 (2H, d, J=6 Hz).

What we claim is:
 1. A compound of the formula: ##STR12## wherein R^(A)is --SCN or --S.sup.⊕ (R³)₂, whereinR³ is lower alkyl, and R^(B) iscarboxy or --COO⁻,or a salt thereof, with the proviso that when R^(A) is--SCN, then R^(B) is carboxy and that when R^(A) is --S.sup.⊕ (R³)₂,then R^(B) is --COO.sup.⊖.
 2. A compound of the formula: ##STR13##wherein R³ is lower alkyl,or a salt thereof.
 3. A compound of theformula: ##STR14## or a salt thereof.
 4. The compound of claim 1,wherein the salt is selected from the group consisting of an organicacid salt, an inorganic acid salt, a metal salt, ammonium salt and anorganic amine salt.
 5. The compound of claim 2, wherein the salt isselected from the group consisting of an organic acid salt, an inorganicacid salt, a metal salt, ammonium salt and an organic amine salt.
 6. Thecompound of claim 3, wherein the salt is selected from the groupconsisting of an organic acid salt, an inorganic acid salt, a metalsalt, ammonium salt and an organic amine salt.